The present invention relates to the recombinant DNA technology. The present invention also relates to the mass-production of antimicrobial materials from microorganisms and a DNA construct and vector system. Biologically active peptide (antimicrobial peptide hereinafter) has little chance to develop resistance since the antimicrobial peptides show activity by a mechanism that is totally different from that of conventional antibiotics which have a serious problem of developing resistance. Therefore, the antimicrobial peptides have a high industrial applicability in the fields of pharmaceutics and the food industry.
The main obstacle in the industrial use of the antimicrobial peptide, however, is the difficulty in economical mass-production of the antimicrobial peptides. For instance, the production of the antimicrobial peptides by chemical synthesis is not economical. Also, there have been attempts to produce antimicrobial peptides by genetic engineering using microorganisms, in this case, however, the expression levels of the antimicrobial peptides are very low.
U.S. Pat. No. 5,206,154 provides a DNA construct which comprises a polypeptide gene which is capable of suppressing the bactericidal effect of cecropin, and a cecropin gene fused to the polypeptide gene. An example of such polypeptide disclosed in the patent is the araB gene.
U.S. Pat. No. 5,593,866 provides a method for a microbial production of a cationic antimicrobial peptide, wherein the cationic peptides is expressed as a fusion to an anionic peptide to avoid degradation by a bacterial protease.
The present invention provides a DNA construct to mass-produce a antimicrobial peptides. The present invention also provides a DNA construct that can produce and recover antimicrobial peptides effectively from microorganisms.
Also, the present invention provides gene multimers that can increase the efficiency of expression, separation and purification of desired peptides and the construction method of such construct.
Further, the present invention provides an expression vector to mass-produce antimicrobial peptides from microorganisms.
Further, the present invention provides a method to mass-produce antimicrobial peptides form microorganisms.